Objective To investigate effect of low molecular weight heparin (LMWH)on oxidative stress and renal function in elderly patients undergoing gastrectomy of stomach neoplasms. Methods 90 elderly patients undergoing radical surgery were randomly divided into LMWH group (Ⅰ group,n=45)and control group (Ⅱ group,n=45). Patients inⅠ group received a subcutaneous injection of LMWH(100 u/kg,1/d)from the time before anesthesia induction to postoperative 5 days,and patients in Ⅱ group were given equal volume of saline. Venous blood samples were collected to determine level of malondialdehyde(MDA),aldose reductase(AR)activity level,superoxide dismutase(SOD)activity level,catalase (CAT)activity level,creatinine(Cr)and blood urea nitrogen(BUN)before anesthesia induction(T0 ),at the end of the surgery(T1 ),on postoperative 1 day(T2),3 days(T3)and 5 days(T4)respectively;urine specimen were also collected to measured albumin(Alb)and N-acetyl-beta-D-amino glycosidase enzymes(NAG)at the same time points. Results There were obvious statistical differences in MDA,AR,SOD and CAT at different time points by intra-group comparison (P<0.05 ). MDA and AR rose first and then fell in two groups,and both reached the peak at T2 ,whereas SOD and CAT had the opposite trends. There were obvious statistical differences in MDA,AR,SOD and CAT at T1 ~T3 (P<0.05 ). MDA and AR inⅠgroup at T1 ,T2 and T3 was lower than those in groupⅡ(P<0.05 ),whereas SOD and CAT inⅠgroup at T1 ,T2 and T3 were significantly higher than those in group Ⅱ(P<0.05). Cr and BUN of both groups were found no significant statistical difference within the group and between groups(P>0.05).Alb/Cr of both groups at T1 and T2 were significantly higher than those at T0 (P<0.05 ),while Alb/Cr in Ⅰ group at T1 was obviously lower than Ⅱgroup(P<0.05 ).NAG/Cr in both groups at T1 ,T2 and T3 were significantly higher than those at T0 (P<0.05 ),while NAG/Cr inⅡ group at T4 was still higher than that at T0 (P<0.05 ),and NAG/Cr inⅠgroup at T1 ~T4 was obviously lower thanⅡgroup (P<0.05 ). MDA and AR in both groups at T1 ,T2 were positively correlated with Alb/Cr and NAG/Cr respectively(P<0.05 ),whereas SOD and CAT in both groups at T1 ,T2 were negatively correlated with Alb/Cr and NAG/Cr respectively(P<0.05 ). Conclusion Oxidative stress reaction resulted from radical surgery of elderly patients is associated with perioperative renal damage. LMWH could reduce oxidative stress in elderly patients,and alleviate the kidney damage,as well as protect the renal function.

Objective To explore the clinico-pathological characteristics of primary breast lymphoma(PBL).Methods The clinical data of 36 cases of PBL were analyzed retrospectively.Results Among the 36 cases of PBL,16 patients presented with stage Ⅰa disease,14 patients with stage IIa disease,2 patients with stageⅡb disease,and 4 patients with stage Ⅳ.All of the patients underwent surgery and chemotherapy,and 20 cases had radiotherapy.Thirty three patients(91.7%) were followed up for 3-193 months,during which time,12 patients died,including 2 patients died of brain metastases,five patients died of bone metastasis,and five patients died of diffuse hepatic and pulmonary metastasis.In the 21 surviving patients,the survival time was 3～192 months,and the median survival time was 43.5 months.The overall 3-and 5-year survival rate was 70.1% and 49.0%,respectively.Conclusions Most PBL are NHL.PBL is diagnosed basically by methods of pathology and immunohistochemistry,and operation combined with chemotherapy and/or radiotherapy is the best treatment method.

Aim To investigate the up-regulation of miR-22 through Wnt pathway inhibits the proliferation,migration and invasion in human gastric MGC803 cells induced by diallyl disulfide(DADS).Methods The effects of proliferation,migration,and invasion of gastric cancer cells were evaluated by MTT,wound-healing and invasion assays.Online prediction software was applied to search the target gene of miR-22.Luciferase report gene assay was used to assess the target genes Wnt-1 of miR-22.The expressions of Wnt-1,β-catenin and TCF-4 were tested by qRT-PCR and Western blot,respectively.Results MTT showed that DADS and miR-22 notably decreased the proliferation compared with control group(P<0.05).Wound-healing assay showed that DADS and miR-22 could significantly inhibit the migration of MGC803 cells compared with the control group, especially in miR-22+DADS(P<0.05). Invasion assay showed that DADS and miR-22 could markedly inhibit the invasion of MGC803 cells compared with the control group, especially in miR-22+DADS(P<0.05). Online prediction software to search the target gene exhibited that Wnt-1 may be a target gene of miR-22. Luciferase report gene assay disclosed that Wnt-1 was identified as a direct target of miR-22. Qrt-PCR showed that the expression of Wnt-1 Mrna was respectively down-regulated by DADS and miR-22 compared withcontrol group, especially in miR-22+DADS(P<0.05). Western blot exhibited that DADS and miR-22 obviously suppressed the expressions of Wnt-1, β-catenin and TCF-4 proteins, especially in miR-22+DADS(P<0.05).Conclusion Up-regulation of miR-22 through Wnt pathway can remarkably suppress the proliferation, migration and invasion in MGC803 cells by DADS.

Animals ; Cell Line, Tumor ; Enoyl-CoA Hydratase ; genetics ; metabolism ; Liver Neoplasms, Experimental ; metabolism ; pathology ; Lymphatic Metastasis ; Mice ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection

ObjectiveTo study the impact of EZH2 silence on the sensitivity of human hepatic multidrug-resistant cancer cells Bel/Fu to 5-Fu.MethodsBel/Fu cells were cultured in vitro; EZH2 siRNA was used to interfere EZH2 expression; RT-PCR and Western blot was used to detect the efficiency of interference.MTT assay was used to detect the cellular growth inhibitory rate; Annexin V-FITC/PI double staining was used to detect the apoptosis rate of cells ; Flow cytometry was to analyze cell cycle ; Western blot analysis was used to detect the expression of multidrug resistance-associated protein MDR1 after silencing EZH2.The experiment set up four groups:control group,5-Fu treatment group,EZH2 siRNA treatment group,5-Fu combined with EZH2 siRNA treatment group. ResultsThe expression of EZH2 was greatly decreased after 24 h in the combined group,the apoptotic inhibitory rate by MTT was 43.17％ ± 3.81％,higher than other three groups; the apoptotic rate in the combined group by Flow cytometry was 30.4％ ± 1.77％,markedly higher than other three groups.The cell cycle of the combined group detected by Flow cytometry was 69.16％ ±2.31％ of cells in the combined group at G1 phase,the percentage was higher than other three group,30.76％ ± 1.29％ at S,G2 and M phases,lower than other three groups,indicating the cell cycle was blocked at G1 phase.MDR1 protein level in the combined group was lower than other groups.ConclusionsSilencing EZH2 strengths the sensitivity of Bel/Fu cells to 5-Fu,probably by a mechanism decreasing the expression of MDR1.

Objective To study the inhibitory effects of δ-opioid receptor activation in serumdeprivation induced apoptosis of human liver cells and the proposed protein kinase C(PKC)pathway mechanism.Methods MTT assay was used to detect the survival rate of human liver cells in vitro and Annexin V-FITC/PI double staining was used to detect the cell apoptosis rate.Flow cytometry was used to analyze cell cycle,RT PCR used to analyze the PKC mRNA and Western Blot analysis was used for detecting the protein expression of PKC and Caspase-3.Results After serum-deprivation for 48h of cultured human liver cells in vitro,significant liver cell apoptosis occurred.The apoptosis was suppressed by δ-opioid receptor activation,which manifested as a slower rate of apoptosis,decreased expression of Caspase-3and increased expression of PKC.After GF109203X was added,the inhibitory effects of DADLE decreased markedly.Conclusion Activation of δ-opioid receptor on the membrane of human liver cells has inhibitory effects on serum-deprivation induced apoptosis of liver cells.The underlying mechanism may be associated with PKC pathway activation.

Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2％ trypsin and 0.133％ collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1％ fibronectin and cultured in endothelial cell-specialized medium supplemented with 10％ fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.

Aim The effects of antibiotics and anticoagulants on mouse peritonaeum were ob-served to explore the factor of the peritoneal dialysis related sclerosing peritoni-tis. Methods The experimental models of peritoneal dialysis were established in miceby infusing different kind of drugs to the peritoneal cavity and the changes of the peri-toneal membrane for each drug at different time were observed by the autopsy and lightmicroscope for several weeks. Results Amikacin, Cefradine, Zinacef, Ciprofloxacin,Heparin and Urokinase could induce sclerosing changes of peritoneal membrane such asloss of peritoneal mesothelum infiltration of inflammatory cells and of proliferation fibrecell.These changes were irreversible after the drugs were stoped.Conclusion Thedrugs commonly used in peritoneal dialysis may in different degree result in peritonealsclerosis.

Objective:To compare the modified Qjng Long Bai Wei needling method and ordinary acupuncture method in the effects of improving the levels of interleukin (IL)-1β,IL-6 and tumor necrosis factor (TNF)-α in the treatment of knee osteoarthritis (KOA),and to determine the advantage of the modified Qing Long Bai Wei needling method for KOA.Methods:One hundred KOA patients were randomized into a treatment group and a control group by using the random number table,with 50 cases in each group.The treatment group was intervened by the modified Qing Long Bai Wei needling method,and the control group was given ordinary acupuncture.The two groups were observed before and after the treatment to determine the changes in the levels of IL-1β,IL-6 and TNF-α in synovial fluid,and the clinical efficacies were compared between the two groups.Results:The total effective rate and clinical recovery rate were 97.9％ and 52.1％ respectively in the treatment group,versus 85.1％ and 25.5％ in the control group,and the between-group differences were statistically significant (both P＜0.01).After the treatment,the levels of Ib1β,IL-6 and TNF-cα in synovial fluid changed significantly in both groups (all P＜0.01);there were significant differences in the levels of IL-1β,IL-6 and TNF-α in synovial fluid between the two groups (all P＜0.01).Conclusion:The modified Qjng Long Bai Wei needling is an effective method for KOA and it can significantly improve the levels of IL-1β,IL-6 and TNF-α in synovial fluid.

OBJECTIVE:To prepare a three-pulse drug release system of nimodipine and study its drug release.METHODS:Solid dispersion technique and dry-coating method were respectively applied to prepare immediate-release mini-tablets and Pulsatile mini-tablets with lag-time of 4 h or 8 h.Then those mini-tablets were filled into capsule to obtain a three-pulse drug release system and subjected to in vitro dissolution test.DSC was employed to determine drug status in solid dispersion.RESULTS:Immediate-release mini-tablets were released more than 95% in 30 min.Pulsatile mini-tablets were released less than 10% in 4 h or 8 h of lag-time period.After lag-time period,pulsatile mini-tablets were released completely in 3 h.The whole pulsatile drug release system achieved three times of drug release at 5 min,4 h,8 h,respectively.Nimodipine kept amorphous form and were delivered into carrier evenly.CONCLUSION:A three-pulse drug release system of nimodipine has been prepared successfully.